| Infectious Hematopoietic Necrosis Virus (IHNV) belongs to the Novirhabdovirus genus, which is an enveloped virus, and possesses a external glycoprotein G inserted into the viral membrane responding for attachment to the membrane of host cell, and help virus get entry into the host cell. An about 1.5kb full length sequence of IHNV-G gene was obtained by RT-PCR amplification. The amplified fragment was cloned into prokaryotic expression system pMAL-c2X vector and then was transformed into Rossetta-gami2 (DE3) splysS. Recombinants were examined with restriction enzyme digestion and further confirmed the interest insert by sequencing pMAL-c2X-G plasmid. SDS-PAGE revealed that the expressed fusion protein is produced by inducing with IPTG, and its molecular weight is around 99kD, which is the prospective size corresponding to the predicted value. In this study, pMAL-c2X was used to reduce hydrophobic property and cytotoxicity of G protein, so the full length of G protein has been expressed successfully and the protein has been confirmed that is soluble. The success of expressing the IHNV G protein is a good starting point for investigating the structure and functions of this protein, and further more to explore the mechanism of the viral entry to cell, and design anti-virus agent or vaccine.Multilplication of IHNV in EPC was investigated and purificated through ultrahigh-speed Centrifugalization. The IHNV particles of high purity can be seen by TEM (transmission electron microscope). The diameter is 110—120nm. The samples of purification can be labled by quantum dot to trace the dynamical process of the viral entry into cell.
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